JP2009531417A - Malates and (3S, 5S) -7- [3-amino-5-methyl-piperidinyl] -1-cyclopropyl-1,4-dihydro-8-methoxy-4-oxo-3-quinolinecarboxylic acid Polymorphs of - Google Patents

Abstract

  The present invention relates to (3S, 5S) -7- [3-amino-5-methylpiperidinyl] -1-cyclopropyl-1,4-dihydro-8-methoxy-4-oxo-3-quinolinecarboxylic acid. It relates to malates and their polymorphs. The invention further relates to pharmaceutical compositions comprising the described salts and polymorphs.

Description

  The present invention relates to (3S, 5S) -7- [3-amino-5-methylpiperidinyl] -1-cyclopropyl-1,4-dihydro-8-methoxy-4-oxo-3-quinolinecarboxylic acid. The present invention relates to malates and various polymorphic forms of malates and pharmaceutical compositions thereof.

  Antibacterial quinolone compounds (3S, 5S) -7- [3-amino-5-methylpiperidinyl] -1-cyclopropyl-1,4-dihydro-8-methoxy-4-oxo-3- Quinoline carboxylic acid and (3S, 5R) -7- [3-amino-5-methyl-piperidinyl] -1-cyclopropyl-1,4-dihydro-8-methoxy-4-oxo-3-quinoline carboxylic acid Is disclosed in US Pat. No. 6,329,391, which is incorporated herein by reference in its entirety. Synthesis of various quinolone compounds is described in US Pat. No. 6,329,391, US Pat. No. 6,803,469, B.I. Ledoussal et al., “Non-6-Fluorosubstituted Quinolone Antibiotics: Structure and Activity” (J. Med Chem., 35, 198-200, 1992), V. Cecchetti et al., "Studies on 6-aminoquinolines: Synthesis and antibacterial evaluation of 6-amino-8-methylquinolines" (J. Med. Chem., 39, 436-445, 1996), V. . Cecchetti et al., “6-Desfluoro-8-methylquinolines effective as novel lead compounds in antibacterial chemotherapy” (J. Med. Chem, 39, 4952-4957, 1996), etc. It has been reported in the literature.

  The above compounds are useful for treating bacterial infections. However, it is not known what salt forms result in suitable preparations for the production of pharmaceutically acceptable compositions. Accordingly, there is a need in the art to produce useful salt forms and polymorphs of these antimicrobial compounds.

  In one aspect, the invention provides:

  (3S, 5S) -7- [3-Amino-5-methyl-piperidinyl] -1-cyclopropyl-1,4-dihydro-8-methoxy-4-oxo-3-quinolinecarboxylic acid (hereinafter referred to as Compound I) (See also intermediate (23) in section D of the best mode for carrying out the invention.)).

  In one aspect, the invention relates to polymorphic malates of Compound I, wherein from about 0% to about 5% by weight of water is present.

  In another aspect, the invention relates to a polymorphic salt of Compound I, wherein from about 1% to about 5% by weight of water is present.

  In another aspect, the invention relates to a polymorphic salt of Compound I, wherein from about 0% to about 2% by weight of water is present.

  In another aspect, the invention relates to a polymorphic salt of Compound I having an X-ray diffraction pattern characterized by substantially matching the pattern of FIG.

  In another aspect, the invention relates to a polymorphic salt of Compound I having an X-ray diffraction pattern characterized by substantially matching the pattern of FIG.

  In another aspect, the invention relates to a polymorphic salt of Compound I having an X-ray diffraction pattern characterized by substantially matching the pattern of FIG.

In another aspect, the invention relates to a polymorphic salt of Compound I having a solid state ¹³ C NMR spectrum characterized by substantially matching the pattern of FIG.

In another aspect, the invention relates to a polymorphic salt of Compound I having a solid state ¹³ C NMR spectrum characterized by substantially matching the pattern of FIG.

In another aspect, the invention relates to a polymorphic salt of Compound I having a solid state ¹³ C NMR spectrum characterized by substantially matching the pattern of FIG.

In another aspect, the invention relates to a polymorphic salt of Compound I having a solid state ¹³ C NMR spectrum characterized by substantially matching the pattern of FIG.

In another aspect, the invention relates to a polymorphic salt of Compound I having a solid state ¹³ C NMR spectrum characterized by substantially matching the pattern of FIG.

  In another aspect, the invention relates to a polymorphic salt of Compound I having an infrared spectrum characterized by substantially matching the pattern of FIG.

  In another aspect, the invention relates to a polymorphic salt of Compound I having an infrared spectrum characterized by substantially matching the pattern of FIG.

  In another aspect, the invention relates to a polymorphic salt of Compound I having an infrared spectrum characterized by substantially matching the pattern of FIG.

  In another aspect, the invention relates to a polymorphic salt of Compound I having an infrared spectrum characterized by substantially matching the pattern of FIG.

  In another aspect, the invention relates to a polymorphic salt of Compound I having an infrared spectrum characterized by substantially matching the pattern of FIG.

  In another aspect, the present invention relates to polymorphic salts of Compound I having characteristic X-ray diffraction peaks at about 10.7 degrees, about 11.98 degrees, and about 12.5 degrees (2θ).

  In another aspect, the invention relates to polymorphic salts of Compound I having characteristic X-ray diffraction peaks at about 9.3 degrees, about 12.1 degrees, and about 22.6 degrees (2θ).

  In another aspect, the invention relates to polymorphic salts of Compound I having characteristic X-ray diffraction peaks at about 9.5 degrees, about 11.7 degrees, and about 12.3 degrees (2θ).

  In another aspect, the present invention relates to D, L-malate hemihydrate, D-malate hydrate, L-malate hydrate, D-malate anhydride, and L-malic acid. It relates to polymorphic salts selected from the group consisting of salt anhydrides.

  In another aspect, the invention relates to a pharmaceutical composition comprising a safe and effective amount of a polymorph according to any of the above polymorphs, and a pharmaceutically acceptable carrier.

  In another aspect, the present invention identifies a human or other animal in need of treatment or prevention of an infectious disease that treats or prevents the treatment of an infectious disease in a human or other animal when needed. And a method comprising administering to a human or other animal a safe and effective amount of the compound of claim 1.

  Herein, various malates and different polymorphs of malates are described. Selection of pharmaceutically acceptable salts with desirable properties (eg, solubility, stability, ease of formulation) requires evaluation of a number of salts and the resulting polymorphs (“drug salts, Handbook of Pharmaceutical Salts, Properties, Selection and Use "(PH Stahl, CG Wermuth, Wiley-VCH, Zurich, 2002) )checking).

  Solids exist in either amorphous or crystalline form. In the crystalline form, the molecules are arranged at three-dimensional lattice sites. When a compound crystallizes out of solution or slurry, it crystallizes with a different spatial lattice arrangement, which is referred to as “polymorphism”, with different crystal forms individually referred to as “polymorphs”. It is a property. Different polymorphic forms of a given substance can interact with each other in terms of one or more physical properties such as solubility and dissolution rate, true density, crystal form, compaction behavior, flowability, and / or solid stability. It can be different.

Crystallization Crystallization on a production scale is accomplished by manipulating the solution to exceed the solubility limit of the compound of interest. This can be achieved by various methods, for example by dissolving the compound at a relatively high temperature and then cooling the solution below the saturation limit. Alternatively, the liquid volume may be reduced by boiling, ambient pressure evaporation, vacuum drying or some other means. The solubility of the compound of interest may be reduced by adding an antisolvent or a solvent or mixture of such solvents that exhibits reduced solubility of the compound. Another option may be to adjust the pH to reduce solubility. For a detailed description of crystallization, see "Crystallization" (3rd edition, written by JW Mullens, Butterworth-Heineman Ltd, 1993, ISBN0750611294). That.

  If salt formation is desired simultaneously with crystallization, the addition of a suitable acid or base results in direct crystallization of the desired salt (if the salt is less soluble than the starting material in the reaction medium). Similarly, if the synthesis reaction in the medium is complete and the final desired form is less soluble than the reactants, direct crystallization of the final product may be possible.

  Crystallization optimization can include seeding the crystallization medium with the desired form of crystals. In addition, many crystallization processes use a combination of the above approaches. The compound of interest is dissolved in the solvent at an elevated temperature, followed by the addition of the antisolvent while controlling the system to an appropriate amount slightly below the saturation level. At this point, seeds of the desired form may be added and the system is allowed to cool and crystallize, leaving the seeds intact.

Pharmaceutical Formulations and Methods of Use The present invention also provides methods of treating or preventing infectious diseases in humans or other animal subjects by administering to the subject a safe and effective amount of a salt or polymorph. . As used herein, an “infectious disease” is a disease characterized by the presence of a microbial infection. A preferred method of the invention is for the treatment of bacterial infections. Such infectious diseases include (eg) central nervous system infections, outer ear infections, middle ear infections (eg acute otitis media), dural sinus infections, eye infections, oral infections (eg dental , Gum and mucous membrane infection), upper respiratory tract infection, lower respiratory tract infection (including pneumonia), genitourinary infection, gastrointestinal infection, gynecological infection, sepsis, rot, peritonitis, osteoarthritis infection , Skin and skin tissue infections, bacterial endocarditis, burns, surgical antibacterial prophylaxis, and antibacterial prophylaxis of postoperative or immunosuppressed patients (eg, patients receiving cancer chemotherapy or organ transplant patients) Can be mentioned.

The salts or polymorphs of the present invention may be administered to treat or prevent various microbial diseases. The pharmaceutical composition is
(A) a safe and effective amount of a salt or polymorph of the invention, and (b) a pharmaceutically acceptable carrier.

  As used herein, the term “treatment” means that administration of a compound of the invention alleviates a disease or disorder in a host. Thus, the term “treatment” specifically refers to prevention of disease development, disease inhibition, and / or disease alleviation in the host, especially when the host is susceptible to the disease but has not yet been diagnosed with the disease. Includes improvements. Where the method of the invention relates to disease prevention, the term “prevention” should be understood that the disease state need not be completely prevented. (See Webster's Ninth Collegiate Dictionary.) Rather, as used herein, the term prevention allows the compound of the present invention to be administered before the disease develops. Including the ability of one skilled in the art to identify populations susceptible to disease. The term does not mean that the medical condition has been completely avoided. The compounds determined by the screening method of the present invention may be administered together with other compounds.

  The safety and therapeutic efficacy of the determined compounds may be determined by standard procedures using in vitro or in vivo techniques. Compounds that exhibit sufficient therapeutic indices may be preferred, but in some cases, compounds with insufficient therapeutic indices are also useful. Data obtained from in vitro and in vivo toxicological and pharmacological techniques may be used to determine dose ranges. The effects of the compounds can be further evaluated in either animal models or patient clinical trials.

  A “safe and effective amount” of a compound of the present invention is effective in inhibiting microbial growth at the site of infection to be treated by the host and has acceptable side effects (eg, toxic, inflammatory, or allergic reactions). It is the amount you have. The specific “safe and effective amount” is the specific condition being treated, the patient's health, the duration of the treatment, the nature of the combination therapy (if any), the specific dosage form used, the recruitment It can vary depending on factors such as the formulation and the desired usage for the composition.

  As used herein, “pharmaceutically acceptable carrier” refers to solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like that are compatible with the administration of the drug. It is intended to include. The use of such media and agents for pharmaceutically active substances is known in the art. Except where any conventional medium or agent is incompatible with the active compound, such medium may be used in the compositions of the present invention. Supplementary compounds may also be incorporated into the compositions. A pharmaceutical composition of the invention is formulated to be compatible with its intended route of administration. Examples of routes of administration include parenteral (eg, intravenous, intradermal, subcutaneous, intramuscular), oral, inhalation, transdermal (topical), transmucosal, and rectal administration. Solutions or suspensions used for parenteral, intradermal, or subcutaneous application include the following components, water for injection, saline solution, fixed oil, polyethylene glycol, glycerin, propylene glycol, or other synthetic solvents Aseptic diluents such as; antibacterial agents such as benzyl alcohol or methyl paraben; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid; buffering agents such as acetate, citrate, or phosphate And tonicity adjusting agents such as sodium chloride or dextrose. The pH can be adjusted with suitable acids or bases. The parenteral preparation can be enclosed in ampoules, disposable syringes or multiple use vials made of glass or plastic.

  Pharmaceutical compositions suitable for injectable use include sterile aqueous solutions (where water soluble) or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersion. For intravenous administration, suitable carriers include physiological saline, Cremophor EL ™ (BASF, Parsippany, NJ), or phosphate buffered saline (PBS). The composition may be sterile and fluid as long as it can be easily injected. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (eg, glycerol, propylene glycol, and polyethylene glycol), and suitable mixtures thereof. Fluidity may be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. Prevention of microbial growth can be achieved with various antibacterial and antifungal agents, such as parabens, chlorobutanol, phenol, ascorbic acid, thimerosal. Examples of isotonic agents include saccharides, polyhydric alcohols such as mannitol and sorbitol, and sodium chloride. Prolonged absorption of injectable compositions can be brought about by including in the composition an agent that delays absorption, for example, aluminum monostearate and gelatin.

  Sterile injectable solutions can be prepared by incorporating the compound in the required amount, one of the above-listed components, or a combination, in a suitable solvent followed by filter sterilization. The dispersion medium may be prepared by incorporating the compound into a sterile excipient that may contain a basic dispersion medium and other ingredients. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred method of preparation is to obtain a powder of any additional desired ingredients in addition to the compound from those pre-sterilized filtered solutions, vacuum dried and frozen Drying is mentioned.

  The oral composition may include an inert diluent or an edible carrier. They may be enclosed in gelatin capsules or pressed into tablets. For oral administration, the agent may be placed in an enteric form so that it can pass through the stomach, or it may be further coated or mixed to be released in specific areas of the gastrointestinal tract by known methods. For the purpose of oral therapeutic administration, the compound may be incorporated with excipients and used in the form of tablets, troches, or capsules. The oral composition may also be prepared using a fluid carrier for use as a mouthwash, in which case the compound in the fluid carrier is applied to the oral cavity and gargles before being exhaled. Or swallowed. Pharmaceutically compatible binding agents, and / or adjuvant materials can be included as part of the composition. Tablets, pills, capsules, lozenges, etc. are the following ingredients: binders such as microcrystalline cellulose, gum tragacanth, or gelatin; excipients such as starch or lactose; alginic acid, Primogel ™, or Disintegrants such as corn starch; lubricants such as magnesium stearate; glidants such as colloidal silicon dioxide; sweeteners such as sucrose or saccharin; or such as peppermint, methyl salicylate, or orange flavor A flavoring agent; or may contain any compound of similar nature.

  For administration by inhalation, the compounds may be supplied in the form of an aerosol spray from pressured container or dispenser which contains a suitable propellant, eg, a gas such as carbon dioxide, or a nebulizer.

  Systemic administration can also be by transmucosal or transdermal means. For transmucosal or transdermal administration, penetrants appropriate to the barrier to be passed may be used in the formulation. Such penetrants are generally known in the art and include, for example, for transmucosal administration, clearing agents, bile salts, and fusidic acid derivatives. Transmucosal administration may be accomplished using spray nasal sprays or suppositories. For transdermal administration, the compounds may be formulated into ointments, salves, gels, or creams as is generally known in the art.

  The compounds may also be prepared in the form of suppositories (eg, with conventional suppository bases such as cocoa butter and other glycerides) or retention enemas for rectal delivery.

  In one embodiment, the compounds are prepared with carriers that prevent the compound from being rapidly excreted from the body, such as controlled release formulations, including implants and microencapsulated delivery systems. Biodegradable and biocompatible polymers may be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid. Methods for preparing such formulations will be apparent to those skilled in the art. Liposomal suspensions can also be used as pharmaceutically acceptable carriers.

  It may be advantageous to formulate oral or parenteral compositions in dosage unit form for ease of administration and uniformity of dosage. As used herein, “dosage unit form” refers to a physically discrete unit that is suitable as a single dosage for the subject to be treated, each unit associated with a pharmaceutical carrier. Contains a predetermined quantity of compound calculated to produce the desired therapeutic effect. The specifications for dosage unit forms of the invention may be dictated by the characteristics of the compound and the particular therapeutic effect to be achieved, as well as limitations inherent in the art of preparing such compounds for the treatment of animals. It can also depend on them.

Example 1: (3S, 5S) -7- [3-Amino-5-methyl-piperidinyl] -1-cyclopropyl-1,4-dihydro-8-methoxy-4-oxo-3-quinolinecarbon Synthesis of acid and its malate A. Synthesis of (3S, 5S)-(5-methyl-piperidin-3-yl) -carbamic acid-tert-butyl ester (8):

(2S) -1- (1,1-dimethylethyl) -5-oxo-1,2-pyrrolidinedicarboxylic acid-2-methyl ester, (2). A 50 liter reaction vessel is charged with compound (1) (5.50 kilograms, 42.60 moles) and methanol (27 liters) and cooled to 10-15 ° C. Thionyl chloride (10.11 kilograms, 2.0 equivalents) is added from the addition funnel over 65 minutes and externally cooled to maintain the temperature below 30 ° C. The resulting solution was stirred at 25 ° C. + 5 ° C. for 1.0 hour, after which methanol was distilled off under reduced pressure. The resulting thick oil is azeotroped with ethyl acetate (3 times, 2.5 liters each) to remove residual methanol. The residue was dissolved in ethyl acetate (27.4 liters), charged into a 50 liter reaction vessel and neutralized by adding triethylamine (3.6 kilograms) from the addition funnel over 30 minutes. The neutralization temperature was maintained below 30 ° C. by external cooling. The resulting triethylamine hydrochloride suspension was removed by filtration and the clear mother liquor was charged to a 50 liter reaction vessel with DMAP (0.53 kilograms). Di-tert-butyl dicarbonate (8.43 kilograms) was added from an addition funnel heated with hot water over 30 minutes with external cooling to maintain the temperature at about 20-30 ° C. The reaction completed after 1 hour is confirmed by TLC analysis. The organic phase is washed with ice-cold 1N HCl (2 times 7.5 liters), saturated sodium bicarbonate solution (1 time, 7.5 liters) and dried over magnesium sulfate. The mixture was filtered through a vacuum filter and the ethyl acetate was removed under reduced pressure to give a crystalline slurry triturated with MTBE (10.0 liters), filtered and intermediate (2) as a white solid (5.45 kilograms, 52.4%). Calculated for analysis C ₁₁ H ₁₇ NO ₅ : found: C, 54.5; H, 6.96; N, 5.80. HRMS (High Resolution Mass Spectrometry) (ESI ⁺ (electrospray positive ion mode)) predicted value for C ₁₁ H ₁₈ NO ₅ , [M + H] 244.1185. Found: 244.1174; ¹ H NMR (CDCl ₃ , 500 MHz): δ = 4.54 (dd, J = 3.1, 9.5 Hz, 1H), 3.7 (s, 3H), 2.58- 2.50 (m, 1H), 2.41 (ddd, 1H, J = 17.6, 9.5, 3.7), 2.30-2.23 (m, 1H), 1.98-1 .93 (m, 1H), 1.40 (s, 9H); ¹³ C NMR (CDCl ₃ , 125.70 MHz) δ 173.3, 171.9, 149.2, 83.5, 58.8, 52. 5, 31.1, 27.9, 21.5; Mp70.2 ° C.

(2S, 4E) -1- (1,1-dimethylethyl) -4-[(dimethylamino) methylene] -5-oxo-1,2-pyrrolidinedicarboxylic acid-2-methyl ester (3). In a 50 liter reaction vessel, intermediate (2) (7.25 kilograms, 28.8 moles), DME (6.31 kilograms), and Bredereck's Reagent (7.7 kilograms, 44.2 moles). Fill. The solution is stirred and heated to 75 ° C. ± 5 ° C. for at least 3 hours. The progress of the reaction is monitored by HPLC. The reaction is cooled to 0 ° C. ± 5 ° C. over 1 hour, during which time a precipitate is formed. The mixture is held at 0 ° C. ± 5 ° C. for 1 hour, filtered through a vacuum filter, and the product is dried in a vacuum oven at 30 ° C. ± 5 ° C. for at least 30 hours to give intermediate (3) as white crystals As a crystalline solid (6.93 kilograms, 77.9%). Calculated for analysis C ₁₄ H ₂₂ N ₂ O ₅ : C, 56.4; H, 7.43; N, 9.39. Found: C, 56.4; H, 7.32; N, 9.48. HRMS (High Resolution Mass Spectrometry) (ESI ⁺ (electrospray positive ion mode)) predicted value for C ₁₄ H ₂₂ N ₂ O ₅ , [M + H] 299.1607. Found 299.1613; ¹ H NMR (CDCl ₃ , 499.8 MHz) δ = 7.11 (s, 1H) 4.54 (dd, 1H, J = 10.8, 3.6), 3.74 ( s, 3H), 3.28-3.19 (m, 1H), 3.00 (s, 6H), 2.97-2.85 (m, 1H), 1.48 (s, 9H); ¹³ C NMR (CDCl ₃ , 125.7 MHz) δ = 172.6, 169.5, 150.5, 146.5, 90.8, 82.2, 56.0, 52.3, 42.0, 28. 1, 26.3. Mp 127.9 ° C.

(2S, 4S) -1- (1,1-dimethylethyl) -4-methyl-5-oxo-1,2-pyrrolidinedicarboxylic acid-2-methyl ester (4). A 37.9 liter (10 gallon) Pfaudler reaction vessel was inerted with nitrogen, ESCAT142, 5% palladium powder on carbon (50% moisture, 0.58 kg, moisture weight), intermediate (3) ( 1.89 kilograms, 6.33 moles) and isopropanol (22.4 kilograms) are charged. The reaction mixture is stirred at 45 ° C. under a hydrogen atmosphere of 0.31 MPa (45-psi) for 18 hours. The reaction mixture is then cooled to room temperature and filtered through a Celite (0.51 kilogram) bed in a vacuum filter to remove the catalyst. The mother liquor is evaporated under reduced pressure to give a thick oil that crystallizes on standing to give 4 (1.69 kilograms, 100%) as a 93: 7 diastereomeric mixture. A sample of the product mixture is purified by preparative HPLC to obtain analytical data material. analysis. Calcd for _{C 12 H 19 NO 5: C} , 56.0; H, 7.44; N, 5.44. Found C, 55.8; H, 7.31; N, 5.44; MS (mass spectrometry) (ESI ⁺ (electrospray positive ion mode)) predicted value for C ₁₂ H ₁₉ NO ₅ , [M + H] 258.1342. Found 258.1321; ¹ H NMR (CDCl ₃ , 499.8 MHz) δ = 4.44 (m, 1H), 3.72 (s, 3H), 2.60-2.48 (m, 2H), 1.59 to 1.54 (m, 1H), 1.43 (s, 9H), 1.20 (d, j = 6.8 Hz, 3H); ¹³ C NMR (CDCl ₃ , 125.7 MHz) δ = 175.7, 172.1, 149.5, 83.6, 57.4, 52.5, 37.5, 29.8, 27.9, 16.2. Mp 89.9 ° C.

(1S, 3S)-(4-Hydroxyl-1-hydroxymethyl-3-methyl-butyl) -carbamic acid tert-butyl ester (5). A 50 liter reaction vessel is charged with intermediate (4) (3.02 kilograms, 11.7 moles), absolute ethanol (8.22 kilograms), and MTBE (14.81 kilograms). The solution is stirred, cooled to 0 ° C. ± 5 ° C., and sodium borohydride (1.36 kilograms, 35.9 mol) is added in portions to keep the reaction temperature at 0 ° C. ± 5 ° C. . Slight bubbling is observed. The reaction mixture is warmed to 10 ° C. ± 5 ° C. and calcium chloride dihydrate (2.65 kilograms) is added in small portions over 1 hour to maintain the reaction temperature at 10 ° C. ± 5 ° C. To be. The reaction is warmed to 20 ° C. ± 5 ° C. over 1 hour and stirred for an additional 12 hours at 20 ° C. ± 5 ° C. The reaction is cooled to −5 ° C. ± 5 ° C. and ice-cold 2N HCl (26.9 kilograms) is added at a rate that maintains the reaction temperature at 0 ° C. ± 5 ° C. Stirring is stopped and the phases are allowed to separate. Remove the lower aqueous phase (pH = 1). A reaction vessel is charged with aqueous saturated sodium bicarbonate (15.6 kilograms) over 5 minutes. Stirring is stopped and the phases are allowed to separate. Remove the lower aqueous phase (pH = 8). A reaction vessel is charged with magnesium sulfate (2.5 kilograms) and stirred for at least 10 minutes. Filter the mixture through a vacuum filter and condense under reduced pressure to give intermediate (5) (1.80 kilograms, 66%). analysis. Calcd for _{C 11 H 23 NO 4: C} , 56.6; H, 9.94; N, 6.00. Found C, 56.0; H, 9.68; N, 5.96. HRMS (High Resolution Mass Spectrometry) (ESI ⁺ (electrospray positive ion mode)) predicted value for C ₁₁ H ₂₄ NO ₄ , [M + H] 234.1705. Found 234.1703; ¹ HNMR (CDCl ₃ , 500 MHz) δ = 6.34 (d, J = 8.9 Hz, 1H, NH), 4.51 (t, J = 5.8, 5.3 Hz, 1H , NHCHCH ₂ OH), 4.34 (t, J = 5.3, 5.3 Hz, 1H, CH ₃ CHCH ₂ OH), 3.46-3.45, (m, 1H, NHCH), 3.28 (dd , J = 10.6, 5.3 Hz, NHCHCHHOH), 3.21 (dd, J = 10.2, 5.8 Hz, 1H, CH ₃ CHCHHOH), 3.16 (dd, J = 10.2, 6 .2 Hz, 1 H, NHCHCHHOH), 3.12 (dd, J = 10.6, 7.1 Hz, 1 H, CH ₃ CHCHHOH), 1.53 to 1.50 (m, 1 H, CH ₃ CHCHHOH), 1. 35 (s, 9H, O ( CH 3) 3, 1.30 ( dd, J = 13.9, 10.2, 3.7 Hz, 1H, NHCHCHHCH), 1.14 (dddd, J = 13.6, 10.2, 3.4 Hz, 1H, NHCHCHHCH), 0.80 ( d, J = 6.6 Hz, 3H, CH ₃ ); ¹³ C NMR (CDCl ₃ , 125.7 MHz) δ 156.1, 77.9, 50.8, 65.1, 67.6, 65.1, 35 .6, 32.8, 29.0, 17.1, Mp 92.1 ° C.

  (2S, 4S) -Methanesulfonic acid-2-tert-butoxycarbonylamino-5-methanesulfonyloxy-4-methyl-pentyl ester (6). A 50 liter reaction vessel is charged with a solution of intermediate (5) (5.1 kilograms) in isopropyl acetate (i-PrOAc) (11.8 kilograms) followed by an additional 7.9 kilograms of i-PrOAc. . Cool the reaction to 15 ° C. ± 5 ° C. and add triethylamine (TEA) (7.8 kilograms) while maintaining the set temperature. The reaction vessel is further cooled to 0 ° C. ± 5 ° C. and methanesulfonyl chloride (MsCl) (6.6 kilograms) is added to the reaction solution while maintaining the set temperature. The reaction is stirred for 2-3 hours and monitored for completion by HPLC or TLC. The reaction is quenched by the addition of saturated aqueous bicarbonate and the resulting isolated organic phase is washed well with 10% cold triethylamine, cold HCl, saturated cold bicarbonate, and finally saturated brine. To wash. The organic phase is dried, filtered and concentrated under reduced pressure at less than 55 ° C. ± 5 ° C. until a solid / liquid slurry containing intermediate (6) is obtained. The slurry is used crude in subsequent reactions without further characterization.

  (3S, 5S)-(1-Benzyl-5-methyl-piperidin-3-yl) -carbamic acid tert-butyl ester (7). A 50 liter reaction vessel is charged with 9.1 kilograms of neat benzylamine. The reaction vessel was brought to 55 ° C., and a solution of intermediate (6) (8.2 kg) in 1,2-dimethoxyethane (DME) (14.1 kg) was added to the reaction vessel while maintaining a temperature of 60 ° C. ± 5 ° C. Add to. After the addition of this solution is complete, the reaction is stirred at 60 ° C. ± 5 ° C. for several hours and monitored for completion by TLC or HPLC. The reaction is cooled to ambient temperature and volatiles (DME) are removed under reduced pressure by rotary evaporation. The residue was diluted with 11.7 kilograms of 15% (v / v) ethyl acetate / hexane solution and treated with 18.7 kilograms of 20% (wt) aqueous potassium carbonate with stirring. By precipitation, a triphasic mixture is obtained. Remove the bottom aqueous phase and separate the intermediate phase. The upper organic phase is collected and saved for combination with the extract from further extraction. The separated intermediate phase is again extracted twice with 11.7 kilogram parts of 15% (volume / volume) ethyl acetate / hexane solution, each time combining the extract with the original organic phase. The combined organic extracts are transferred to a rotary evaporator and the solvent is removed under reduced pressure until an oily residue remains. The residue is then purified by large scale preparative chromatography to give the purified intermediate (7) as an oil.

(3S, 5S)-(5-Methyl-piperidin-3-yl) -carbamic acid-tert-butyl ester (8). A 40 liter pressure vessel is filled with 0.6 kg, 50% moisture, solid palladium on carbon (E101, 10 wt%) under a stream of nitrogen. Next, a solution containing 3.2 kg of intermediate (7) in 13.7 kg of absolute ethanol is charged into a reaction vessel in a nitrogen atmosphere. The reaction vessel may be purged with nitrogen and then pressurized to 0.31 MPa (45 psi) with hydrogen. The reaction is then heated to 45 ° C. while maintaining the hydrogen pressure at 0.31 MPa (45 psi). Monitor by TLC or LC until reaction is complete. Cool the reaction to ambient temperature, vent and purge with nitrogen. Filter the contents of the reaction vessel through a celite bed and wash the solids with 2.8 kilograms of absolute ethanol. The filtrate is concentrated by rotary evaporation under reduced pressure until a waxy solid is obtained, yielding intermediate (8). TLC R _f (silica F ₂₅₄ , 70:30 (volume / volume) ethyl acetate-hexane, KMnO ₄ stain) = 0.12; ¹ H NMR (300 MHz, CDCl ₃ ) δ 5.31 (br s, 1H), 3 .80 to 3.68 (m, 1H), 2.92 (d, J = 11.4 Hz, 1H), 2.77 (AB quarter, J _AB = 12.0 Hz, Δv = 50.2 Hz, 2H), 2.19 (t, J = 10.7 Hz, 1H), 1.82-1.68 (m, 2H), 1.54 (brs, 1H), 1.43 (s, 9H), 1.25 1.15 (m, 1H), 0.83 (d, J = 6.6 Hz, 3H); ¹³ C NMR (75 MHz, CDCl ₃ ) δ 155.3, 78.9, 54.3, 50.8, 45 .3, 37.9, 28.4, 27.1, 19.2; MS (mass spectrometry) (ESI + ( Direct b spray positive ion mode)) m / z215 (M + H), 429 (2M + H).

  B. Synthesis of 1-cyclopropyl-7-fluoro-8-methoxy-4-oxo-1,4-dihydro-quinoline-3-carboxylic acid (19):

Intermediate (12): A reaction vessel was charged with a solution of intermediate (11) (1.2 kilograms, 7.7 moles, 1.0 equivalent) in anhydrous toluene (12 liters) followed by ethylene glycol (1.8 liters). 15.7 moles, 4.2 equivalents) and solid p-toluenesulfonic acid (120 grams, 10% by weight). The reaction mixture is stirred at ambient temperature for at least 30 minutes and then heated to reflux until the reaction completion can be confirmed by TLC analysis (15% EtOAc / hexanes volume / volume) and the water / toluene azeotrope. Are collected with a Dean-Stark trap device. When the reaction is complete, the reaction is cooled to ambient temperature and poured into aqueous sodium bicarbonate (6 liters). The organic toluene layer was removed and washed with saturated sodium bicarbonate solution (6 liters), distilled water (twice, 6 liters), and saturated aqueous brine (6 liters). The organic phase was removed, dried over MgSO ₄ , filtered and evaporated under reduced pressure to give intermediate (12) as an oil (1.3 kilogram, 86%). The material is used in subsequent reaction steps without further purification.

  Intermediate (13): A reaction vessel was charged with a solution of intermediate (12) (1.2 kilograms, 6.0 mol, 1.0 eq) in anhydrous tetrahydrofuran (12 liters) and n-butyllithium (2 in hexane). 0.5M, 2.6 liters, 6.6 moles, 1.1 equiv.) At −40 ° C. while maintaining this temperature during the addition. The reaction is stirred for at least 1 hour at −40 ° C. and trimethyl borate (0.9 liter, 7.8 mol, 1.3 eq) is added to the mixture while maintaining the temperature below −40 ° C. The reaction mixture is stirred at −40 ° C. for at least 1 h until reaction completion can be confirmed by TLC analysis (30% EtOAc / hexanes vol / vol). The reaction mixture is warmed in portions to −30 ° C. and acetic acid (3 L) is added slowly. When the addition is complete, water (0.5 liter) is added to the reaction and the mixture is quickly warmed to ambient temperature and stirred overnight. The organic solvent is removed from the reaction by distillation at 45 ° C. under reduced pressure. To the reaction residue, 3-4 volumes of water (6 liters) and 30% hydrogen peroxide (0.7 liters, 1.0 equivalent) are slowly added while cooling at ambient temperature to control the exotherm. To do. The reaction mixture is stirred at ambient temperature for at least 1 hour until reaction completion can be confirmed by TLC (15% EtOAc / hexanes vol / vol). The reaction mixture is cooled to 0-5 ° C. and excess peroxide is quenched by the addition of 10% aqueous sodium bisulfite (2 liters). The mixture is inspected to ensure that no peroxide is present and the reaction is acidified by adding 6N HCl (aq) (1.2 liters). The reaction is stirred until the hydrolysis reaction is complete as confirmed by TLC or NMR analysis. The resulting solid is collected by suction filtration to give intermediate (13) as a yellow solid (1.0 kilogram, 79%).

  Intermediate (14): A reaction vessel is charged with Intermediate (13) (0.53 kilograms, 3.0 moles, 1.0 equivalent) and dissolved in dry toluene (2.7 kilograms, 3.1 liters). Let To this solution is added dimethyl sulfate (0.49 kilograms, 3.9 moles, 1.30 equivalents) followed by solid potassium carbonate (0.58 kilograms, 4.2 moles, 1.4 equivalents). The reaction mixture is heated to reflux and held for at least 1 hour until reaction completion can be confirmed by HPLC. During this time, intense outgassing is observed. The reaction is then cooled to ambient temperature and diluted with distilled water (3.2 liters) along with 30% NaOH (aq) (0.13 kilograms, 0.33 equivalents). The aqueous phase was separated and the residual toluene layer was extracted more than once with distilled water (3.2 liters) mixed with 30% NaOH (aq) (0.13 kilograms, 0.33 equivalents), each time the aqueous phase Remove. The upper organic phase is concentrated under reduced pressure (less than 10 kPa (100 mbar)) by distillation at about 40 ° C. until a concentrated toluene solution is obtained. The resulting solution was cooled to ambient temperature, quality and yield were confirmed by HPLC, and the following synthesis step was performed without further purification (the theoretical yield of intermediate (14) was 0.56 kilograms). Is presumed to be).

  Intermediate (15a, b): Sodium hydride (0.26 kg, 6.6 mol, 2.20 eq) as a dispersion in 60 wt% mineral oil with 1.8 kg (2.1 liters) anhydrous toluene At the same time, the reaction vessel is filled. To this mixture is added diethyl carbonate (0.85 kilogram, 7.2 mol, 2.4 eq) as a reaction mixture and heated to 90 ° C. for over 1 hour. While maintaining a temperature of 95 ° C. ± 5 ° C., a toluene solution (˜1.0 equivalent) of intermediate (14) obtained in the previous step is added to the reaction. Gas evolution can be observed during this addition. After the addition is complete, the reaction is stirred for at least 30 minutes or until complete by HPLC analysis. When the reaction is complete, the mixture is cooled to ambient temperature and diluted with 10% by weight aqueous sulfuric acid (3.8 kilograms, 3.9 moles, 1.3 equivalents) with stirring. The phases are separated and the lower aqueous phase is removed. The residual organic phase is concentrated under reduced pressure (less than 10 kPa (100 mbar)) at about 40 ° C. until a concentrated toluene solution is obtained. The resulting solution is cooled to ambient temperature and subjected to the next synthetic step without further purification (theoretical yield of intermediate (15a, b) is estimated to be 0.85 kilograms).

  Intermediate (16a, b; 17a, b): Fill a reaction vessel with a toluene solution of intermediate (15a, b) (0.85 kilogram, ~ 3.0 mol, ~ 1.0 equivalent) obtained in the previous step. did. Next, dimethylformamide-dimethylacetal (0.54 kilograms, 4.5 moles, 1.5 equivalents) is added to the reaction vessel and the resulting solution is heated to reflux (up to 95-105 ° C.). The bottom boiling solvent (methanol from the reaction) is distilled off while maintaining the temperature above 90 ° C. Continue heating for at least 1 hour or until reaction completion is confirmed by HPLC analysis. Upon completion of the reaction, the reaction containing a mixture of intermediates (16a, b) was cooled to ambient temperature and toluene (1) with cyclopropylamine (0.21 kilograms, 3.6 moles, 1.2 equivalents). .8 kilograms, 2.1 liters) is added to the reaction. The reaction is stirred at ambient temperature for at least 30 minutes until HPLC analysis confirms reaction completion. Once the reaction is complete, the reaction is diluted with 10 wt% aqueous sulfuric acid (2.9 kilograms, 3.0 moles, 1.0 equiv) with stirring and then the phases are separated. The aqueous phase is removed and the organic phase is concentrated by distillation at about 40 ° C. under reduced pressure (less than 10 kPa (100 mbar)). When the desired concentration is reached, the solution is cooled to ambient temperature and the following synthesis step is performed without further purification of the toluene solution containing the mixture of intermediates (17a, b) (intermediate (17a, b)). Is estimated to be ~ 1.1 kilograms).

  Intermediate (18): A solution of a mixture of intermediates (17a, b) (˜4.7 kilograms, −3.0 moles) is charged to the reaction vessel at ambient temperature. To the reaction vessel is added N, O-bis (trimethylsilyl) acetamide (0.61 kilograms, 3.0 moles, 1.0 equiv) and the reaction is refluxed for at least 30 minutes or until complete by HPLC analysis. Heat to temperature (up to 105-115 ° C). If the reaction is not complete, additional N, 0-bis (trimethylsilyl) acetamide (0.18 kilogram, 0.9 mol, 0.3 eq) is added to complete the reaction. When the reaction is complete, the reaction is cooled to less than 40 ° C. and the organic solvent is removed by distillation under reduced pressure (less than 10 kPa (100 mbar)) at about 40 ° C. until a precipitate is formed. The reaction is cooled to ambient temperature and the precipitated solid is separated by suction filtration and washed twice with distilled water (1.8 liter once, 0.9 liter once). The solid is dried to give intermediate (18) as a white solid (0.76 kilogram, 82%). The material is used in the next reaction step without further purification.

Intermediate (19): A reaction vessel was charged with solid intermediate (18) (0.76 kilograms, ~ 2.5 moles, ~ 1.0 equivalents) at ambient temperature followed by ethanol (5.3 kilograms). 6.8 liters) and 32% by weight aqueous hydrochloric acid (1.1 kilograms, 10 moles). The reaction mixture is brought to reflux temperature (76-80 ° C.), during which time the mixture initially becomes homogeneous and later becomes heterogeneous. The mixture is heated at reflux for at least 5 hours or until reaction completion is confirmed by TLC analysis (15% EtOAc / hexanes vol / vol). When the reaction is complete, the reaction is cooled to 0 ° C. ± 5 ° C. and the precipitated solid is separated by filtration and washed with distilled water (1.7 kilograms) followed by ethanol (1.7 kilograms). The separated solid is dried to give intermediate (19) as a white solid (0.65 kilogram, ˜95%). ¹ H NMR (CDCl ₃ , 300 MHz) δ (ppm): 14.58 (s, 1H), 8.9 (s, 1H), 8.25 (m, 1H), 7.35 (m, 1H), 4.35 (m, 1H), 4.08 (s, 3H), 1.3 (m, 2H), 1.1 (m, 2H). ¹⁹ F NMR (CDCl ₃ + CFCl ₃ , 292 MHz) δ (ppm): −119. HPLC: 99.5% (area).

  C. Synthesis of boron ester chelate (20) of 1-cyclopropyl-7-fluoro-8-methoxy-4-oxo-1,4-dihydro-quinoline-3-carboxylic acid

A reaction vessel is charged with boron oxide (2.0 kilograms, 29 moles) followed by dilution with glacial acetic acid (8.1 liters, 142 moles) and acetic anhydride (16.2 liters, 171 moles). The resulting mixture is heated to reflux temperature for at least 2 hours. The reaction contents are cooled to 40 ° C. and solid 7-fluoroquinolinic acid intermediate (19) (14.2 kilograms, 51 mol) is added to the reaction mixture. The mixture is heated again to reflux temperature for at least 6 hours. The reaction progress is monitored by HPLC and NMR. The mixture is cooled to about 90 ° C. and toluene (45 liters) is added to the reaction. Further, the reaction is cooled to 50 ° C. and 3-butyl methyl ether (19 liters) is added to the reaction mixture to precipitate the product. The mixture is then cooled to 20 ° C. and the solid product 19 is separated by filtration. The separated solids are then washed with 3-butyl methyl ether (26 liters) prior to drying in a vacuum (6.67 kPa (50 torr)) oven at 40 ° C. The product yield obtained from intermediate (20) of this reaction is 86.4%. Raman (cm ^-1 ): 3084.7, 3022.3, 2930.8, 1709.2, 1620.8, 1548.5, 1468.0, 1397.7, 1368.3, 1338.5, 1201.5 955.3, 653.9, 580.7, 552.8, 384.0, 305.8. NMR (CDCl ₃ , 300 MHz) δ (ppm): 9.22 (s, 1H), 8.38-8.33 (m, 1H), 7.54 (t, J = 9.8 Hz, 1H), 4 .38-4.35 (m, 1H), 4.13 (s, 3H), 2.04 (s, 6H), 1.42-1.38 (m, 2H), 1.34-1.29 (M, 2H). TLC (Whatman MKC18F silica, 60 Å, 200 micrometers), mobile phase: 1: 1 (volume / volume) CH ₃ CN: 0.5N NaCl (aq), UV (254/366 nanometers) visualization; R _f = 0.4-0.5.

  D. 1-cyclopropyl-7-fluoro-8-methoxy-4-oxo-1,4-dihydro-quinoline-3-carboxylic acid (20) (3S, 5S)-(5-methyl-piperidin-3-yl) -Coupling to carbamic acid tert-butyl ester (8) and (3S, 5S) -7- [3-amino-5-methylpiperidinyl] -1-cyclopropyl-1,4-dihydro-8 Synthesis of malate salt of -methoxy-4-oxo-3-quinolinecarboxylic acid (25):

A reaction vessel was charged with solid intermediate (20) (4.4 kilograms, 10.9 moles) followed by a solution of triethylamine (TEA) (2.1 liters, 14.8 moles) and acetonitrile (33.5 Dilute at room temperature with piperidine side chain intermediate (8) (2.1 kilograms, 9.8 moles) in liters, 15.7 liters per kilogram). The resulting mixture is warmed to about 50 ° C. until the reaction is complete. The reaction progress is monitored by HPLC or reverse phase TLC. When the reaction is complete, the reaction is cooled to about 35 ° C. and the volume of the reaction is reduced to about half by distilling acetonitrile under reduced pressure between 0 kPa (0 torr) and 53.3 kPa (400 torr). The reaction vessel is then charged with 28.2 kilograms of 3.0N NaOH (aq) solution and the temperature is raised to about 40 ° C. Continue vacuum distillation until no distillate is observed for 1-4 hours or longer. The reaction is then cooled to room temperature and the hydrolysis reaction is monitored by HPLC or reverse phase TLC. Once the reaction is complete, the reaction mixture is neutralized to pH 6-8 by adding ~ 4-5 kilograms of glacial acetic acid. The reaction vessel is then charged with 12.7 kilograms (9.6 liters) of dichloromethane as an extraction solvent, the mixture is stirred, the phases are separated, and the organic dichloromethane phase is removed. Repeat the extraction process two more times using 12.7 kilograms (9.6 liters) of dichloromethane, each time collecting the lower organic phase. Discard the aqueous phase and integrate the organic extracts into one reaction vessel. The contents of the reaction vessel are heated to 40 ° C. and the volume of the reaction is reduced by about half by distillation. The reaction vessel is then charged with 20.2 kilograms of 6.0 N HCl (aq) solution, the temperature is adjusted to about 35 ° C. and stirred for at least 12 hours to initiate the Boc deprotection reaction. The reaction is monitored by HPLC or reverse phase TLC. When the reaction is complete, stirring is stopped and the phases are allowed to separate. The lower organic phase is removed and set aside. The reaction vessel is then charged with 12.7 kilograms (9.6 liters) of dichloromethane as an extraction solvent, the mixture is stirred, the phases are separated, and the organic dichloromethane phase is removed. Integrate organic extracts and discard. The remaining aqueous phase is diluted with 18.3 kilograms of distilled water and the temperature is raised to about 50 ° C. Vacuum distillation (13.3 kPa (100 torr) to 53.3 kPa (400 torr)) is performed to remove residual dichloromethane from the reaction. Next, the pH of the reaction is adjusted to 7.8-8.1 using approximately 9.42 kilograms of 3.0N NaOH (aq) solution while maintaining the temperature of the reaction below 65 ° C. . The reaction is cooled to 50 ° C. and the precipitated solid is allowed to stand for at least 1 hour before the mixture is cooled to room temperature. The solid is separated by suction filtration and washed twice with 5.2 kilogram portions of distilled water. Dry the solids by aspiration for at least 12 hours, then dry at 55 ° C. in a convection oven for an additional 12 hours. The yield obtained for intermediate (23) of this example is 3.2 kilograms (79%). A reaction vessel is charged with 3.2 kilograms of solid intermediate (23) and the solid is suspended in 25.6 kilograms of 95% ethanol as solvent. To the reaction vessel, 1.1 kilograms of solid D, L-malic acid (24) is then added and the mixture is heated to reflux temperature (˜80 ° C.). Distilled water (˜5.7 liters) is added to the reaction until the reaction is complete, and 0.2 kilograms of active charcoal is added. The reaction mixture is purified through a filter, cooled to 45 ° C. and held for at least 2 hours for crystallization to occur. The reaction mixture is further cooled to 5 ° C. and the suspended solid is separated by suction filtration. The solid is then washed with 6.6 kilograms of 95% ethanol and sucked dry under reduced pressure for at least 4 hours. The solids are then further dried in a convection oven at 45 ° C. for at least 12 hours to yield 3.1 kilograms of intermediate (24) (70%). NMR (D _{2 0,} 300 MHz) δ (ppm): 8.54 (s, 1H), 7.37 (d, J = 9.0 Hz, 1H), 7.05 (d, J = 9.0 Hz) 1H), 4.23 to 4.18 (m, 1H), 4.10 to 3.89 (m, 1H), 3.66 (brs, 1H), 3.58 (s, 3H), 3. 45 (d, J = 9.0 Hz, 1H), 3.34 (d, J = 9.3 Hz, 1H), 3.16 (d, J = 12.9 Hz, 1H), 2.65 (dd , J = 16.1, Hz, 4.1 Hz, 1H), 2.64 to 2.53 (m, 1H), 2.46 (dd, J = 16.1 Hz, 8.0 Hz, 1H), 2 .06 (br s, 1H), 1.87 (d, J = 14.4 Hz, 1H), 1.58 to 1.45 (m, 1H), 1.15 to 0.95 (m, 2H) , 0.91 (d, J = 6 3 Hz, 3H), 0.85~0.78 (m, 2H). TLC (Whatman MKC18F silica, 60 Angstroms, 200 micrometers), mobile phase: 1: 1 (volume / volume) CH ₃ CN: 0.5N NaCl (aq), UV (254/366 nanometers) visualization. HPLC: Gradient elution with _{H 2 O, 88% H 2} O / formic acid to 20% H ₂ O / formic acid containing mobile phase of 0.1% formic acid / acetonitrile and 0.1% formic acid, Zorbax (Zorbax) SB-C8 4 .6 mm x 150 mm column. Part No. 883975.906, 1.5 ml / min / speed, 20 min / operation time, 292 nm, detector model G1314A, S / N / JP72003849, Quat pump model G1311A, S / N / US72102299, automatic sampler model G1313A , S / N · DE149918139, Degasser model G1322A, S / N · JP73007229; estimated retention time of intermediate (19): 13.0 minutes; estimated retention time of intermediate (20): 11.6 minutes; intermediate ( Approximate retention time of 21): 16.3 minutes; Approximate retention time of intermediate (22): 18.2 minutes; Approximate retention time of intermediate (23): 8.6 minutes; Approximate retention time of compound (25) : 8.6 minutes.

Example 2 Salt Preparation and Salt Form Evaluation A salt screen is performed on 100 mg of free base. The separated salt form is evaluated by NMR, elemental analysis, TG-DTA, XRD, and HPLC. Table 1 describes the physical and chemical characteristics of these salt forms. As shown therein, malates can provide a balance between desired solubility, stability, and ease of separation. In addition, the use of malates may be useful for chiral purification. Furthermore, D, L-malate, D-malate or L-malate can provide different advantages depending on the nature of the chiral impurity being removed. Hydrated forms may provide better wettability and solid stability in addition to excellent ease of separation. Use of the anhydrous form may clearly improve the solubility and dissolution rate. Thus, the malates of Compound I exhibit certain advantages, including ease of separation, reduced hygroscopicity, greater solubility in water, greater stability, and ease of formulation.

Example 3: Preparation of D, L-malate hemihydrate of Compound I Synthesis of D, L-malate salt of Compound I from the free base: 10 grams of the free base of Compound I and 1 equivalent of D, L-malic acid were heated in 105 mL of 95% ethanol to reflux (about 78 ° C). Add 15 mL of water while maintaining the temperature near 78 ° C. Continue stirring and heating until completely dissolved. Additional water may be added to ensure complete dissolution. With stirring, the solution is slowly cooled (over at least 3 hours) to room temperature to initiate crystallization. If an oily or waxy mass (or phase other than hemihydrate) precipitates, the solution is reheated to completely dissolve the precipitate and cool more slowly. The crystalline solid is then filtered and washed with a small amount of 95% ethanol. The crystals are dried at ambient pressure, room temperature, and relative humidity between 25% and 75%.

  B. Crystallization of existing malate salt of Compound I: 10 grams of D, L-malate salt of Compound I is heated in 105 mL of 95% ethanol and refluxed (about 78 ° C.). Add 15 mL of water while maintaining the temperature near 78 ° C. Continue stirring and heating until the salt is completely dissolved. Additional water may be added to ensure complete dissolution. With stirring, the solution is slowly cooled (over at least about 3 hours) to room temperature to initiate crystallization. If an oily or waxy mass (or phase other than hemihydrate) precipitates, the solution is reheated to completely dissolve the precipitate and cool more slowly. The crystalline solid is then filtered and washed with a small amount of 95% ethanol.

  The crystals are dried at ambient pressure, room temperature, and relative humidity between 25% and 75%.

Example 4 Preparation of D-malate hydrate of Compound I Synthesis of D-malate salt of Compound I from the free base: 10 grams of the free base of Compound I and 1 equivalent of D-malic acid are heated in 75 mL of 95% ethanol to reflux (about 78 ° C.). Add 25 mL of water while maintaining the temperature near 78 ° C. Continue stirring and heating until completely dissolved. Additional water may be added to ensure complete dissolution. With stirring, the solution is slowly cooled (over at least 3 hours) to room temperature to initiate crystallization. If an oily or waxy mass (or non-hydrated phase) precipitates, the solution is reheated to completely dissolve the precipitate and cool more slowly. The crystalline solid is then filtered and washed with a small amount of 95% ethanol. The crystals are dried at ambient pressure, room temperature, and relative humidity between 25% and 75%.

  B. Crystallization of existing D-malate salt of Compound I: 10 grams of D-malate salt of Compound I is heated in 75 mL of 95% ethanol and refluxed (about 78 ° C.). Add 25 mL of water while maintaining the temperature near 78 ° C. Continue stirring and heating until the salt is completely dissolved. Additional water may be added to ensure complete dissolution. With stirring, the solution is slowly cooled (over at least 3 hours) to room temperature to initiate crystallization. If an oily or waxy mass (or non-hydrated phase) precipitates, the solution is reheated to completely dissolve the precipitate and cool more slowly. The crystalline solid is then filtered and washed with a small amount of 95% ethanol. The crystals are dried at ambient pressure, room temperature, and relative humidity between 25% and 75%.

Example 5: Preparation of L-malate hydrate of Compound I Synthesis of L-malate salt of Compound I from the free base: 10 grams of the free base of Compound I and 1 equivalent of L-malic acid are heated in 75 mL of 95% ethanol to reflux (about 78 ° C.). Add 25 mL of water while maintaining the temperature near 78 ° C. Continue stirring and heating until completely dissolved. Additional water may be added to ensure complete dissolution. With stirring, the solution is slowly cooled (over at least 3 hours) to room temperature to initiate crystallization. If an oily or waxy mass (or non-hydrated phase) precipitates, the solution is reheated to completely dissolve the precipitate and cool more slowly. The crystalline solid is then filtered and washed with a small amount of 95% ethanol. The crystals are dried at ambient pressure, room temperature, and relative humidity between 25% and 75%.

  B. Crystallization of existing L-malate salt of Compound I: 10 grams of L-malate salt of Compound I is heated in 75 mL of 95% ethanol to reflux (about 78 ° C.). Add 25 mL of water while maintaining the temperature near 78 ° C. Continue stirring and heating until the salt is completely dissolved. Additional water may be added to ensure complete dissolution. With stirring, the solution is slowly cooled (over at least 3 hours) to room temperature to initiate crystallization. If an oily or waxy mass (or non-hydrated phase) precipitates, the solution is reheated to completely dissolve the precipitate and cool more slowly. The crystalline solid is then filtered and washed with a small amount of 95% ethanol. The crystals are dried at ambient pressure, room temperature, and relative humidity between 25% and 75%.

Example 6: Preparation of Compound I D-malate Anhydride 280 mg of Compound I D-malate hemihydrate is heated to 70 ° C in 5 mL of dry methanol. Continue heating and stirring until the salt is completely dissolved. The solution is then slowly cooled to room temperature with stirring (cooling over at least about 3 hours). The crystals are filtered and dried under a dry nitrogen purge to protect the sample from moisture during the drying process.

Example 7: Preparation of Compound I L-malate Anhydride 200 mg of Compound I L-malate hemihydrate is heated to 70 ° C in 2 mL of dry methanol. Continue heating and stirring until the salt is completely dissolved. Cool the solution very slowly to room temperature. The solution can be stirred for an additional period of time until crystallization occurs, or the solution can be evaporated with dry nitrogen to cause more rapid crystallization, resulting in material from water absorption during the crystallization and separation process. Protect.

Example 8: Analysis of Polymorphs Various polymorphs that may be obtained using the methods described above can be further characterized using the techniques described below.

  The moisture content is determined by thermogravimetric analysis (TG). Water analysis is performed using Perkin Elmer TGA-7. Samples (5-12 mg) are placed in an open aluminum sample frying pan under dry nitrogen and run at a scan rate of 5 ° C./min.

  The observed moisture content for intact hemihydrate and hydrate ranges from 1.5% to 3.0%. To reduce water content, hydrates and hemihydrates may be dried, and fully hydrated material spectroscopy and XRD signatures may still be maintained. The moisture content observed for the anhydride ranged from “not detected” to 1.0%.

  X-ray diffraction analysis: X-ray powder diffraction is performed on the sample using a Bruker D5000 X-ray diffractometer. The D5000 is equipped with a 2.2 kW Cu anode X-ray tube, Anton Parr TTK-1 low temperature stage, and a high speed position sensitive detector (PSD). CuK radiation (= 1.5418 Angstroms) is used to obtain a powder pattern. A double foil nickel filter is placed in the x-ray receiving path to remove Kβ-radiation. The material was mounted and analyzed on the front loading sample holder. Scan over a range of 3.5 to 40 (2θ), 0.02 step size, 0.2 second for each step.

Solid state nuclear magnetic resonance (SSNMR) analysis: All data are recorded on a Varian 300 Unity Inova spectrometer equipped with a 7 mm CPMAS probe (spinning at 5 kHz). The 75.4 MHz ¹³ C spectrum is recorded with a cross-polarization magic angle spinning (CP / MAS) TOSS (all sideband suppression) instrument. The sample is not powdered but is filled in a 7 mm silicon nitride rotor as it is.

  Infrared (IR) analysis: Samples are analyzed by the split mull method using a BioRad FTS-3000 FTIR spectrometer (with KBr beam splitter). 16 background and sample scans are obtained at a wavenumber resolution of 4 for each sample. Using an agate mortar and agate pestle, mix approximately 1% of the sample with a suitable admixture (eg, fluorolubes for wave numbers 4000-1350, nujol for wave numbers 1350-450). Sample preparation consisting of: The sample may not be powdered prior to mixing with the admixture. A background scan is obtained by using a corresponding KBr disk against which the kneaded sample is sandwiched for sample analysis.

Example 9: Characteristics of various salt forms 7- [3S-amino-5S-methyl-piperidinyl] -1-cyclopropyl-1,4-dihydro-8-methoxy-4 under practical production conditions -Oxo-3-quinolinecarboxylic acid malates may be formed and separated. Depending on the chiral malate (as a racemic mixture or chirally pure form) for salt formation, optionally 7- [3S-amino-5S-methyl-piperidinyl] -1-cyclopropyl-1,4-dihydro-8-methoxy Chiral purification of -4-oxo-3-quinolinecarboxylic acid may be assisted. As a class, malates are slightly soluble in water (according to US Pharmacopoeia 28 definition) and exhibit favorable chemical stability. The hydrated form shows phase stability for relative humidity below 75% according to dynamic vapor sorption measurements and static humidity chamber studies. Using the same test method, it was shown that the anhydrous form absorbs moisture and is exposed to moisture and converted to the corresponding hydrated form naturally.

D, L-malate hemihydrate of compound I The structure of D, L-malate hemihydrate is finally determined by single crystal X-ray diffraction. The smallest unit of this part is two molecules of 7- [3S-amino-5S-methyl-piperidinyl] -1-cyclopropyl-1,4-dihydro-8-methoxy-4-oxo-3-quinolinecarboxylic acid, It consists of one molecule of D-malic acid, one molecule of L-malic acid and one molecule of water. Hydrate water has channel properties, so the water content changes to some extent depending on the relative humidity.

D-malate hydrate and L-malate hydrate of Compound I D-malate hydrate and L-malate hydrate are rapidly separated from the aqueous solvent system as crystalline solids. Good. In order to carry out the separation without problems, it is necessary to use a chirally pure acid. Similar to D, L-malate hemihydrate, hydrated water is channel-like and the moisture content depends to some extent on relative humidity.

D-malate anhydride and L-malate anhydride of Compound I Neither form of anhydride can be separated by crystallites of sufficient size to produce a high definition X-ray diffraction pattern . Anhydride separation often results in oils or waxes that slowly crystallize into high surface area materials. Anhydrides produce a powder pattern consistent with the nanocrystalline material. The resulting X-ray diffraction pattern has a very small signal and inseparable peaks. Nanocrystalline high surface area anhydrides convert to the corresponding hydrate form upon exposure to moisture.

  Except as otherwise specified, all amounts including amounts, percentages, portions, and proportions are understood to be adjusted by the word “about” and amounts are not intended to indicate significant figures.

  Except as otherwise noted, the articles “a”, “an”, and “the” mean “one or more”.

  All documents cited in “Best Mode for Carrying Out the Invention” are incorporated herein by reference in the relevant part, and any citation of any document shall be regarded as prior art to the present invention. It should not be construed as acceptable. To the extent that any meaning or definition of a term in this document conflicts with any meaning or definition of a term in a document incorporated by reference, the meaning or definition given to that term in this document applies And

  While particular embodiments of the present invention have been illustrated and described, it would be obvious to those skilled in the art that various other changes and modifications can be made without departing from the spirit and scope of the invention. Accordingly, it is intended to cover in the appended claims all such changes and modifications that are within the scope of this invention.

2 shows a representative X-ray diffraction pattern of a polymorphic salt of D, L-malate hemihydrate of Compound I. 2 shows a representative X-ray diffraction pattern of a polymorph salt of D-malate hydrate of Compound I. 2 shows a representative X-ray diffraction pattern of a polymorph salt of L-malate hydrate of Compound I. 2 shows a representative solid state ¹³ C NMR spectrum of a polymorphic salt of the D, L-malate hemihydrate of Compound I. 2 shows a representative solid state ¹³ C NMR spectrum of a polymorph salt of D-malate hydrate of Compound I. 2 shows a representative solid state ¹³ C NMR spectrum of polymorph salt of L-malate hydrate of Compound I. 2 shows a representative solid state ¹³ C NMR spectrum of polymorphic salt of Compound I D-malate anhydride. 2 shows a representative solid state ¹³ C NMR spectrum of polymorphic salt of L-malate anhydride of Compound I. 2 shows a representative infrared spectrum of a polymorphic salt of D, L-malate hemihydrate of Compound I. 2 shows a representative infrared spectrum of a polymorph salt of D-malate hydrate of Compound I. 2 shows a representative infrared spectrum of a polymorph salt of L-malate hydrate of Compound I. 2 shows a representative infrared spectrum of polymorphic salt of D-malate anhydride of Compound I. 2 shows a representative infrared spectrum of polymorph salt of L-malate anhydride of Compound I.

Claims (16)

  Malate of (3S, 5S) -7- [3-amino-5-methylpiperidinyl] -1-cyclopropyl-1,4-dihydro-8-methoxy-4-oxo-3-quinolinecarboxylic acid.   The malate according to claim 1, which is a polymorphic salt containing 0% to 5% by weight of water.   Malate according to claim 2, wherein 1% to 5% by weight of water is present.   Malate according to claim 2, wherein 0% to 2% by weight of water is present.   4. Malate salt according to claim 3, having an X-ray diffraction pattern characterized by substantially matching the pattern of FIG. 1, FIG. 2, or FIG. 4. Malate salt according to claim 3, having a solid state ¹³ C NMR spectrum characterized in that it substantially matches the pattern of FIG. 4, FIG. 5 or FIG. 5. Malate salt according to claim 4, having a solid state ¹³ C NMR spectrum characterized in that it substantially matches the pattern of FIG. 7 or FIG.   The malate of claim 3, having an infrared spectral pattern characterized by substantially matching the pattern of FIG. 9, FIG. 10, or FIG.   Malate according to claim 4, having an infrared spectral pattern characterized in that it substantially matches the pattern of figure 12 or figure 13.   The malate salt of claim 3, having characteristic X-ray diffraction peaks at about 10.7 degrees, about 11.98 degrees, and about 12.5 degrees (2θ).   The malate salt of claim 3, having characteristic X-ray diffraction peaks at about 9.3 degrees, about 12.1 degrees, and about 22.6 degrees (2θ).   The malate salt of claim 3, having characteristic X-ray diffraction peaks at about 9.5 degrees, about 11.7 degrees, and about 12.3 degrees (2θ).   Polymorphs selected from D, L-malate hemihydrate, D-malate hydrate, L-malate hydrate, D-malate anhydride, and L-malate anhydride The malate according to claim 1, wherein the malate is a salt.   D, L- of (3S, 5S) -7- [3-amino-5-methyl-piperidinyl] -1-cyclopropyl-1,4-dihydro-8-methoxy-4-oxo-3-quinolinecarboxylic acid Malate hemihydrate polymorphic salt. a. A safe and effective amount of malate according to any one of claims 1 to 14, and b. A pharmaceutical composition comprising a pharmaceutically acceptable carrier.   15. Use of malate according to any one of claims 1 to 14 in the manufacture of a medicament for use in such treatment in humans or other animals in need of treatment or prevention of infectious diseases.

Images